CA2624949C - Substituted 2-(2,6-dioxopiperidin-3-yl)-phthalimides and -1-oxoisoindolines and method of reducing tnf.alpha. levels - Google Patents

Abstract

(R)- and (S)- isomers of Actimid, 1,3-dioxo-2- (2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, having the following formula:

Description

k SUBSTITUTED 2-(2,6-DIOXOPIPERIDIN-3-YL)-METHOD OF REDUCING TNFa LEVELS
This application is a second divisional of Canadian Patent Application SN

2,261,762 filed July 24, 1997. A first divisional application, Canadian Patent Application SN
2,560,523 was filed on September 22, 2006.
It should be understood that the expression "the present invention" or the like used in this specification encompasses not only the subject matter of this second divisional application but that of the parent application and first divisional application also.

The present invention relates to substituted 2-(2,6-diosopiperidin-3-yl)phthal-imides and substituted 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines, the method of reducing levels of tumor necrosis factor a in a mammal through the administration thereof, and pharmaceutical compositions of such derivatives.

Background of the Invention Tumor necrosis factor a, or TNFa, is a cytokine which is released primarily by mononuclear phagocytes in response to a number immunostimulators. When administered to animals or humans, it causes inflammation, fever, cardiovascular effects, hemorrhage, coagulation, and acute phase responses similar to those seen during acute infections and shock states. Excessive or unregulated TNFa production thus has been implicated in a number of disease conditions. These include endo-toxemia and/or toxic shock syndrome {Tracey et al., Nature 330, 662-664 (1987) and Hinshaw et at., Circ. Shock 30, 279-292 (1990)}; cachexia {Dezube et al., Lancet, 335 (8690), 662 (1990)) and Adult Respiratory Distress Syndrome where TNFa concentration in excess of 12,000 pg/mL have been detected in pulmonary aspirates from ARDS patients (Millar et at., Lancet 2(8665), 712-714 (1989)). Systemic infu-sion of recombinant TNFa also resulted in changes typically seen in ARDS
{Ferrai-Baliviera et al., Arch. Surg. 124(12), 1400-1405 (1989)).

TNFa appears to be involved in bone resorption diseases, including arthritis.
When activated, leukocytes will produce bone-resorption, an activity to which the data suggest TNFa contributes. {Bertolini et al., Nature 319, 516-518 (1986) and Johnson et al., Endocrinology 124(3), 1424-1427 (1989).) TNFa also has been shown to stimulate bone resorption and inhibit bone formation in vitro and in vivo through stimulation of osteoclast formation and activation combined with inhibition of osteoblast function. Although TNFa may be involved in many bone resorption diseases, including arthritis, the most compelling link with disease is the association between production of TNFa'by tumor or host tissues and malignancy associated hypercalcemia {Calci. Tissue Int. (US) 46(Suppl.), S3-10 (1990)). In Graft versus Host Reaction, increased serum TNFa levels have been associated with major complication following acute allogenic bone marrow transplants (Holler et al., Blood, 75(4), 1011-1016 (1990)).

Cerebral malaria is a lethal hyperacute neurological syndrome associated with high blood levels of TNFa and the most severe complication occurring in malaria patients.
Levels of serum TNFa correlated directly with the severity of disease and the prog-nosis in patients with acute malaria attacks {Grau et al., N. Engl. J. Med.
320(24), 1586-1591 (1989)).

Macrophage-induced angiogenesis TNFa is known to be mediated by TNFa.
Leibovich et al. {Nature, 329, 630-632 (1987)) showed TNFa induces in vivo capillary blood vessel formation in the rat cornea and the developing chick chorioallantoic membranes at very low doses and suggest TNFa is a candidate for inducing angiogenesis in inflammation, wound repair, and tumor growth. TNFa production also has been associated with cancerous conditions, particularly induced tumors {Ching et al., Brit. J. Cancer, (1955) 72, 339-343, and Koch, Progress in Medicinal Chemistry, 22, 166-242 (1985)).

TNFa also plays a role in the area of chronic pulmonary inflammatory diseases.
The deposition of silica particles leads to silicosis, a disease of progressive respiratory failure caused by a fibrotic reaction. Antibody to TNFa completely blocked the silica-induced lung fibrosis in mice (Pignet et al., Nature, 344:245-247 (1990)).
High levels of TNFa production (in the serum and in isolated macrophages) have been demonstrated in animal models of silica and asbestos induced fibrosis {Bissonnette et al., Inflammation 13(3), 329-339 (1989)). Alveolar macrophages from pulmonary sarcoidosis patients have also been found to spontaneously release massive quantities of TNFa as compared with macrophages from normal donors {Baughman et al.,J. Lab. Clin. Med. 115(1), 36-42 (1990)).

TNF(x is also implicated in the inflammatory response which follows reperfusion, called reperfusion injury, and is a major cause of tissue damage after loss of blood flow {Vedder et al., PNAS 87, 2643-2646 (1990)). TNFa also alters the properties of endothelial cells and has various pro-coagulant activities, such as producing an r T

increase in tissue factor pro-coagulant activity and suppression of the anticoagulant protein C pathway as well as down-regulating the expression of thrombomodulin {Sherry et al., J. Cell Biol. 107, 1269-1277 (1988)). TNFa has pro-inflammatory activities which together with its early production (during the initial stage of an inflammatory event) make it a likely mediator of tissue injury in several important disorders including but not limited to, myocardial infarction, stroke and circulatory shock. Of specific importance may be TNFa-induced expression of adhesion mole-cules, such as intercellular adhesion molecule (ICAM) or endothelial leukocyte adhesion molecule (ELAM) on endothelial cells {Munro el al., Am. J. Path.
135(1), 121-132 (1989)).

TNFa blockage with monoclonal anti-TNFa antibodies has been shown to be beneficial in rheumatoid arthritis {Elliot et al., Int. J. Pharmac. 1995 17(2), 141-145) and Crohn's disease {von Dullemen et al., Gastroenterology, 1995 109(1), 129-135) Moreover, it now is known that TNFa is a potent activator of retrovirus replication including activation of HIV-1. {Duh et al., Proc. Nat. Acad. Sci. 86, 5974-(1989); Poll et al., Proc. Nat. Acad. Sci. 87, 782-785 (1990); Monto et al., Blood 79, 2670 (1990); Clouse et al., J. Immunol. 142, 431-438 (1989); Poll et al., AIDS
Res.
Hum. Retrovirus, 191-197 (1992)). AIDS results from the infection of T
lymphocytes with Human Immunodeficiency Virus (HIV). At least three types or strains of HIV
have been identified, i.e., HIV-1, HIV-2 and HIV-3. As a consequence of HIV
infection, T-cell mediated immunity is impaired and infected individuals manifest severe opportunistic infections and/or unusual neoplasms. HIV entry into the T
lymphocyte requires T lymphocyte activation. Other viruses, such as HIV-1, HIV-infect T lymphocytes after T cell activation and such virus protein expression and/or replication is mediated or maintained by such T cell activation. Once an activated T
lymphocyte is infected with HIV, the T lymphocyte must continue to be maintained in an activated state to permit HIV gene expression and/or HIV replication.
Cytokines, specifically TNFa, are implicated in activated T-cell mediated HIV protein expression and/or virus replication by playing a role in maintaining T lymphocyte activation.
Therefore, interference with cytokine activity such as by prevention or inhibition of cytokine production, notably TNFa, in an HIV-infected individual assists in limiting the maintenance of T lymphocyte caused by HIV infection.

Monocytes, macrophages, and related cells, such as kupffer and glial cells, also have been implicated in maintenance of the HIV infection. These cells, like T
cells, are targets for viral replication and the level of viral replication is dependent upon the activation state of the cells. (Rosenberg et al., The Immunopathogenesis of HIV
Infection, Advances in Immunology, 57 (1989)). Cytokines, such as TNFa, have been shown to activate HIV replication in monocytes and/or macrophages {Poli et al., Proc. Natl. Acad. Sci., 87, 782-784 (1990)}, therefore, prevention or inhibition of cytokine production or activity aids in limiting HIV progression for T cells.
Addi-tional studies have identified TNFa as a common factor in the activation of HIV in vitro and has provided a clear mechanism of action via a nuclear regulatory protein found in the cytoplasm of cells (Osborn, et al., PNAS 86 2336-2340). This evidence suggests that a reduction of TNFa synthesis may have an antiviral effect in HIV
infections, by reducing the transcription and thus virus production.

AIDS viral replication of latent HIV in T cell and macrophage lines can be induced by TNFa {Folks et al., PNAS 86, 2365-2368 (1989)}. A molecular mechanism for the virus inducing activity is suggested by TNFa's ability to activate a gene regulatory protein (NFKB) found in the cytoplasm of cells, which promotes HIV replication through binding to a viral regulatory gene sequence (LTR) {Osborn et al., PNAS
86, 2336-2340 (1989)). TNFa in AIDS associated cachexia is suggested by elevated serum TNFa and high levels of spontaneous TNFa production in peripheral blood monocytes from patients {Wright et al., J. Immunol. 141(1), 99-104 (1988)).
TNFa has been implicated in various roles with other viral infections, such as the cytomega-lia virus (CMV), influenza virus, adenovirus, and the herpes family of viruses for similar reasons as those noted.

The nuclear factor KB (NFKB) is a pleiotropic transcriptional activator (Lenardo, et al., Cell 1989, 58, 227-29). NFKB has been implicated as a transcriptional activator in a variety of disease and inflammatory states and is thought to regulate cytokine levels including but not limited to TNFa and also to be an activator of HIV
transcription (Dbaibo, et al., J. Biol. Chem. 1993, 17762-66; Duh et al., Proc. Natl. Acad.
Sci.
1989, 86, 5974-78; Bachelerie et al., Nature 1991, 350, 709-12; Boswas et al., J.
Acquired Immune Deficiency Syndrome 1993, 6, 778-786; Suzuki et al., Biochem.
And Biophys. Res. Comm. 1993, 193, 277-83; Suzuki et al., Biochem. And Biophys.
Res Comm. 1992, 189, 1709-15; Suzuki et al., Biochem. Mol. Bio. Int. 1993, 31(4), r T

700; Shakhov et al., Proc. Natl. Acad. Sci. USA 1990, 171, 35-47; and Staal et al., Proc. Natl. Acad. Sci. USA 1990, 87, 9943-47). Thus, inhibition of NFiB
binding can regulate transcription of cytokine gene(s) and through this modulation and other mechanisms be useful in the inhibition of a multitude of disease states. The compounds described herein can inhibit the action of NFiB in the nucleus and thus are useful in the treatment of a variety of diseases including but not limited to rheuma-toid arthritis, rheumatoid spondylitis, osteoarthritis, other arthritic conditions, septic shock, septis, endotoxic shock, graft versus host disease, wasting, Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythrematosis, ENL in leprosy, HIV, AIDS, and opportunistic infections in AIDS. TNFa and NFiB levels are influenced by a reciprocal feedback loop. As noted above, the compounds of the present invention affect the levels of both TNFa and NFKB.

Many cellular functions are mediated by levels of adenosine 3',5'-cyclic monophos-phate (cAMP). Such cellular functions can contribute to inflammatory conditions and diseases including asthma, inflammation, and other conditions (Lowe and Cheng, Drugs of the Future, 17(9), 799-807, 1992). It has been shown that the elevation of cAMP in inflammatory leukocytes inhibits their activation and the subsequent release of inflamma-tory mediators, including TNFa and NFKB. Increased levels of cAMP also leads to the relaxation of airway smooth muscle.

Decreasing TNF(x levels and/or increasing cAMP levels thus constitutes a valuable therapeutic strategy for the treatment of many inflammatory, infectious, immunologi-cal, and malignant diseases. These include but are not restricted to septic shock, sepsis, endotoxic shock, hemodynamic shock and sepsis syndrome, post ischemic reperfusion injury, malaria, mycobacterial infection, meningitis, psoriasis, congestive heart failure, fibrotic disease, cachexia, graft rejection, oncogenic or cancerous conditions, asthma, autoimmune disease, opportunistic infections in AIDS, rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, other arthritic conditions, Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupus erythrematosis, ENL in leprosy, radiation damage, oncogenic conditions, and hyperoxic alveolar injury. Prior efforts directed to the suppression of the effects of TNFa have ranged from the utilization of steroids such as dexamethasone and prednisolone to the use of both polyclonal and monoclonal antibodies {Beutler et al., Science 234, 470-474 (1985);
WO 92/11383}.

Detailed Description The present invention is based on the discovery that certain classes of non-polypeptide compounds more fully described herein decrease the levels of TNFa.

In particular, the invention pertains to (i) compounds of the formula:
R' RZ X O
'N1( NH

in which:

one of X and Y is C=O and the other of X and Y is C=O or CH2;

(1) each of R', R2, R3, and R4, independently of the others, is halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms or (ii) one of R', R2, R3, and R4 is -NHR5 and the remaining of R', R2, R3, and R4 are hydrogen;

R5 is hydrogen or alkyl of 1 to 8 carbon atoms;

R6 is hydrogen, alkyl of 1 to 8 carbon atoms, benzyl, or halo;

provided that R6 is other than hydrogen if X and Y are C=O and (i) each of R', R2, R3, and R4 is fluoro or (ii) one of R', R2, R3, or R4 is amino; and (b) the acid addition salts of said compounds which contain a nitrogen atom capable of being protonated.

A preferred group of compounds are those of Formula I in which each of R', R2, R3, and R4, independently of the others, is halo, alkyl of I to 4 carbon atoms, or alkoxy of I to 4 carbon atoms, and R6 is hydrogen, methyl, ethyl, or propyl. A
second preferred group of compounds are those of Formula I in which one of R', R2, R3, and R4 is -NH2, the remaining of R', R2, R3, and R4 are hydrogen, and R6 is hydrogen, methyl, ethyl, or propyl.

CA 02624949 2008-04-04, Unless otherwise defined, the term alkyl denotes a univalent saturated branched or straight hydrocarbon chain containing from I to 8 carbon atoms. Representative of such alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl. Alkoxy refers to an alkyl group bound to the remainder of the molecule through an ethereal oxygen atom. Representative of such alkoxy groups are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy.
Preferably R', R2, R3, and R4 are chloro, fluoro, methyl or methoxy.

The compounds of Formula I are used, under the supervision of qualified profes-sionals, to inhibit the undesirable effects of TNFa. The compounds can be adminis-tered orally, rectally, or parenterally, alone or in combination with other therapeutic agents including antibiotics, steroids, etc., to a mammal in need of treatment.

The compounds of the present invention also can be used topically in the treatment or prophylaxis of topical disease states mediated or exacerbated by excessive TNFa production, respectively, such as viral infections, such as those caused by the herpes viruses, or viral conjunctivitis, psoriasis, atopic dermatitis, etc.

The compounds also can be used in the veterinary treatment of mammals other than humans in need of prevention or inhibition of TNFa production. TNFa medi-ated diseases for treatment, therapeutically or prophylactically, in animals include dis-ease states such as those noted above, but in particular viral infections.
Examples include feline immunodeficiency virus, equine infectious anaemia virus, caprine arthritis virus, visna virus, and maedi virus, as well as other lentiviruses.

53686-5E(S) - 7a -Thus, in one aspect, the invention provides an optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, having the following structure:

O
N...... 0 NH

or a pharmaceutically acceptable salt thereof.

Thus, in one aspect, the invention provides an optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, having the following structure:

O
\ N O
NH

or a pharmaceutically acceptable salt thereof.

Thus, in one aspect, the invention provides an optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, having the following structure:

O
N...... 0 NH

O O

or a pharmaceutically acceptable salt thereof.

53686-5E(S) - 7b -Thus, in one aspect, the invention provides an optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, having the following structure:

O

O O

or a pharmaceutically acceptable salt thereof.

In another aspect, the invention provides use of a compound as described herein for reducing levels of TNFa in a mammal, inhibiting angiogenesis, or treating inflammatory disease, autoimmune disease or a cancerous condition.

In another aspect, the invention provides a pharmaceutical composition comprising a compound as described herein and a pharmaceutically acceptable diluent or carrier.

In another aspect, the invention provides a pharmaceutical composition comprising a compound as described herein and a pharmaceutically acceptable diluent or carrier for reducing levels of TNFa in a mammal, inhibiting angiogenesis, or treating inflammatory disease, autoimmune disease or a cancerous condition.

According to still another aspect of the present invention, there is provided a commercial package comprising the pharmaceutical composition as described herein together with instructions for use for reducing levels of TNFa in a mammal, inhibiting angiogenesis, or treating inflammatory disease, autoimmune disease or a cancerous condition.

53686-5E(S) - 7c -Compounds in which one of R', R9 is amino and R-' and R , as well as the remainder of R~, R`, R', R4, are hydrogen, as for example, 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline or 1,3-dioxo-2-(2,6-dioxopiper ddin-3-yl)-5-aminoisoindoline are known. See, e.g., Jonsson, Acta Pharma. Succica, 9, 521-542 (1972).

The compounds can be prepared using methods which are known in general. In particular, the compounds can be prepared through the reaction of 2,6-dioxopiperidin-3-ammonium chloride, and a lower alkyl ester of 2-bromomethylben-zoic acid in the presence of an acid acceptor such as dimethylaminopyridine or triethylamine.

R2 COOa&yl CrH3N+ N,H
101 R6 --~ I
R3 CH2Br 0 The substituted benzoate intermediates are known or can be obtained though conventional processes. For example, a lower alkyl ester of an ortho-toluic acid is brominated with N-bromosuccinimide under the influence of light to yield the lower alkyl 2-bromomethylbenzoate.

Alternatively, a dialdehyde is allowed to react with 2,6-dioxopiperidin-3-ammonium chloride:

R2 CHO CrH3N+ ,H
101 + R6 N --~' I

In a further method, a dialdehyde is allowed to react with glutamine and the resulting 2-(1-oxoisoindolin-2-yl)glutaric acid then cyclized to yield a I-oxo-2-(2,6-dioxopiperidin-3-yl)-isoindoline of Formula 1:

r 1 RI
R2 CHO C1fH3N+ COOH
101 + R6 ---'~

R' 0 Finally, an appropriately substituted phthalidimide intermediate is selectively reduced:

R' 0 Amino compounds can be prepared through catalytic hydrogenation of the corresponding nitro compound:

N
O, N CH
Y

IA

The nitro intermediates of Formula IA are known or can be obtained though conventional processes. For example, a nitrophthalic anhydride is allowed to react with a-aminoglutarimide hydrochloride {alternatively named as 2,6-dioxopiperidin-3-ylammonium chloride) in the presence of sodium acetate and glacial acetic acid to yield an intermediate of Formula IA in which X and Y are both C=O.

In a second route, a lower alkyl ester of nitro-ortho-toluic acid is brominated with N-bromosuccinimide under the influence of light to yield a lower alkyl 2-(bromomethyl)nitrobenzoate. This is allowed to react with 2,6-dioxopiperidin-3-ammonium chloride in, for example, dimethylforinamide in the presence of trieth-ylamine to yield an intermediate of Formula 11 in which one of X is C=O and the other is CH2.

Alternatively, if one of R1, R2, R3, and R4 is protected amino, the protecting group can be cleaved to yield the corresponding compound in which one of R1, R2, R3, and R4 is amino. Protecting groups utilized herein denote groups which generally are not found in the final therapeutic compounds but which are intentionally intro-duced at some stage of the synthesis in order to protect groups which otherwise might be altered in the course of chemical manipulations. Such protecting groups are removed at a later stage of the synthesis and compounds bearing such protecting groups thus are of importance primarily as chemical intermediates (although some derivatives also exhibit biological activity). Accordingly the precise structure of the protecting group is not critical. Numerous reactions for the formation and removal of such protecting groups are described in a number of standard works including, for example, "Protective Groups in Organic Chemistry", Plenum Press, London and New York, 1973; Greene, Th. W. "Protective Groups in Organic Synthesis", Wiley, New York, 1981; "The Peptides", Vol. 1, Schroder and Lubke, Academic Press, London and New York, 1965; "Methoden der organischen Chemie", Houben-Weyl, 4th Edition, Vol.15/I, Georg Thieme Verlag, Stuttgart 1974.
An amino group can be protected as an amide utilizing an acyl group which is selectively removable under mild conditions, espe-cially benzyloxycarbonyl, formyl, or a lower alkanoyl group which is branched in I-or a position to the carbonyl group, particularly tertiary alkanoyl such as pivaloyl, a lower alkanoyl group which is substituted in the position a to the carbonyl group, as for example trifluoroacetyl.

-11- _ The compounds of the present invention possess a center of chirality and can exist as optical isomers. Both the racemates of these isomers and the individual isomers themselves, as well as diastereomers when there are two chiral centers, are within the scope of the present invention. The racemates can be used as such or can be separated into their individual isomers mechanically as by chromatography using a chiral adsorbent. Alternatively, the individual isomers can be prepared in chiral form or separated chemically from a mixture by forming salts with a chiral acid, such as the individual enantiomers of 10-camphorsulfonic acid, camphoric acid, a-bromocam-phoric acid, methoxyacetic acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid, and the like, and then freeing one or both of the resolved bases, optionally repeating the process, so as obtain either or both substan-tially free of the other; i.e., in a form having an optical purity of >95%.

The present invention also pertains to the physiologically acceptable non-toxic acid addition salts of the compounds of Formula I. Such salts include those derived from organic and inorganic acids such as, without limitation, hydrochloric acid, hydro-bromic acid, phosphoric acid, sulfuric acid, methanesulphonic acid, acetic acid, tartaric acid, lactic acid, succinic acid, citric acid, malic acid, maleic acid, sorbic acid, aconitic acid, salicylic acid, phthalic acid, embonic acid, enanthic acid, and the like.

Oral dosage forms include tablets, capsules, dragees, and similar shaped, com-pressed pharmaceutical forms containing from I to 100 mg of drug per unit dosage.
Isotonic saline solutions containing from 20 to 1 00 mg/mL can be used for parenteral administration which includes intramuscular, intrathecal, intravenous and intra-arterial routes of administration. Rectal administration can be effected through the use of suppositories formulated from conventional carriers such as cocoa butter.

Pharmaceutical compositions thus comprise one or more compounds of the present invention associated with at least one pharmaceutically acceptable carrier, diluent or excipient. In preparing such compositions, the active ingredients are usually mixed with or diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule or sachet. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material which acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, elix-irs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.
Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidinone, cellulose, water, syrup, and methyl cellulose, the formulations can additionally include lubricat-ing agents such as talc, magnesium stearate and mineral oil, wetting agents, emulsify-ing and suspending agents, preserving agents such as methyl- and propylhydroxyben-zoates, sweetening agents or flavoring agents.

The compositions preferably are formulated in unit dosage form, meaning physi-cally discrete units suitable as a unitary dosage, or a predetermined fraction of a unitary dose to be administered in a single or multiple dosage regimen to human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient. The compositions can be formulated so as to provide an immediate, sustained or delayed release of active ingredient after administration to the patient by employing procedures well known in the art.

Oral dosage forms include tablets, capsules, dragees, and similar shaped, compres-sed pharmaceutical forms containing from I to 100 mg of drug per unit dosage.
Isotonic saline solutions containing from 20 to 100 mg/mL can be used for parenteral administration which includes intramuscular, intrathecal, intravenous and intra-arterial routes of administration. Rectal administration can be effected through the use of suppositories formulated from conventional carriers such as cocoa butter.
Pharmaceutical compositions thus comprise one or more compounds of the present invention associated with at least one pharmaceutically acceptable carrier, diluent or excipient. In preparing such compositions, the active ingredients are usually mixed with or diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule or sachet. When the excipient serves as a diluent, it may be a solid, semi-solid, or liquid material which acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, elix-irs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.
Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidinone, cellulose, T

water, syrup, and methyl cellulose, the formulations can additionally include lubricat-ing agents such as talc, magnesium stearate and mineral oil, wetting agents, emulsify-ing and suspending agents, preserving agents such as methyl- and propylhydroxyben-zoates, sweetening agents or flavoring agents.

The compositions preferably are formulated in unit dosage form, meaning physi-cally discrete units suitable as a unitary dosage, or a predetermined fraction of a unitary dose to be administered in a single or multiple dosage regimen to human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient.

The compositions can be formulated so as to provide an immediate, sustained or delayed release of active ingredient after administration to the patient by employing procedures well known in the art.

The following examples will serve to further typify the nature of this invention but should not be construed as a limitation in the scope thereof, which scope is defined solely by the appended claims.

EXAMPLE I
1,3-Dioxo-2-(2,6-dioxopiperidin-3-yi)-5-aminoisoindoline A mixture of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline (altern-atively named as N-(2,6-dioxopiperidin-3-yl)-4-nitrophthalimide} (1 g, 3.3 mmol) and 10% Pd/C (0.13 g) in 1,4-dioxane (200 mL) was hydrogenated at 50 psi for 6.5 hours.
The catalyst was filtered through CelitVand the filtrate concentrated in vacuo. The residue was crystallized from ethyl acetate (20 mL) to give 0.62 g (69%) of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline {alternatively named as N-(2,6-dioxopiperidin-3-yl)-4-aminophthalimide} as an orange solid. Recrystallization from dioxane/ethyl acetate gave 0. 32 g of yellow solid: mp 318.5-320.5 C; HPLC
(nova Pak C18,15/85 acetonitrile/0.1%H3P04) 3.97 min (98.22%). 'H NMR (DMSO-d6) 8 11.08(s, I H), 7.53-7.50 (d, J=8.3 Hz, I H), 6.94(s, IH). 6.84-6.81(d. J=8.3 Hz,1 H), 6.55(s,2H). 5.05-4.98(m, IH), 2.87-1.99(m, 4H); 13C NMR (DMSO-d6) 6 172.79.170.16, 167.65, 167.14, 155.23, 134.21, 125.22, 116.92, 116.17, 107.05, WO 98/03502 PCr/US97/13375 48.58, 30.97, 22.22; Anal. Calcd for C13HõN304: C, 57.14; H, 4.06; N, 15.38.
Found:
C, 56.52- H, 4.17; N, 14.60.

In a similar fashion from I-oxo-2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline, I-oxo-2-(2,6-dioxopiperidin-3-yl)-4-nitroisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-6-nitroisoindoline, I-oxo-2-(2,6-dioxopiperidin-3-yl)-7-nitroisoindoline, and 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-nitroisoindoline, there is respectively obtained 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, I-oxo-2-(2,6-dioxopiperidin-3-yl)-6-aminoisoindoline, I-oxo-2-(2,6-dioxopiperidin-3-yl)-7-aminoisoindoline, and 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, respectively, upon hydrogenation.

1,3-Dioxo-2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline A mixture of 4-nitrophthalic anhydride (1.7 g, 8.5 mmol), a-aminoglutarimide hydrochloride (1.4 g, 8.5 mmol) and sodium acetate (0.7 g, 8.6 mmol) in glacial acetic acid (30 mL) was heated under reflux for 17 hours. The mixture was concentrated in vacua and the residue was stirred with methylene chloride (40 mL) and water (30 mL). The aqueous layer was separated, extracted with methylene chloride (2x40 mL).
The combined methylene chloride solutions were dried over magnesium sulfate and concentrated in vacuo to give 1.4 g (54%) of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline as a light brown solid. An analytical sample was obtained by recrystallization from methanol: mp 228.5-229.5 C; 'H NMR (DMSO-d6) 6 11.18(s, H), 8.69-8.65(d,d J=1.9 and 8.0 Hz, IH), 8.56(d, J=1.9 Hz, 1H), 8.21(d, H=8.2 Hz, 1H), 5.28(d,d J=5.3 and 12.8 Hz, 1H), 2.93-2.07(m, 4H); 13C NMR (DMSO-d6) S
172.66, 169.47, 165.50, 165.23, 151.69, 135.70, 132.50, 130.05, 124.97, 118.34, 49.46, 30.85, 21.79; Anal. Calcd for C, 3H9N306: C, 51.49; H, 2.99; N, 13.86.
Found:
C, 51.59; H, 3.07; N, 13.73.

1-Oxo-2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline, I-oxo-2-(2,6-dioxopip-eridin-3-yl)-4-nitroisoindoline, 1-oxo-2-(2,6-dioxopiperidin-'I-yl)-6-nitroisoindoline, and 1-oxo-2-(2,6-dioxopiperidin-3-yl)-7-nitroisoindoline can be obtained by allowing 2,6-dioxopiperi din-3-ammonium chloride to react with methyl 2-bromomethyl-5-nitrobenzoate, methyl 2-bromomcthyl-4-nitrobenzoate, methyl 2-bromomethyl-6-~, T

nitrobenzoate, and methyl 2-bromomethyl-7-nitrobenzoate, respectively, in dimethyl-formamide in the presence of triethylamine. The methyl 2-(bromomethyl)nitro-benzoates in turn are obtained from the corresponding methyl esters of nitro-ortho-toluic acids by conventional bromination with N-bromosuccinimide under the influ-ence of light.

1-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetraflu oroisoindoline A mixture of 16.25 g of 2,6-dioxopiperidin-3-ammonium chloride, and 30.1 g of methyl 2-bromomethyl-3,4,5,6-tetrafluorobcnzoate, and 12.5 g of triethylamine in 100 mL of dimethylformamide is stirred at room temperature for 15 hours. The mixture is then concentrated in vacuo and the residue mixed with methylene chloride and water. The aqueous layer is separated and back-extracted with methylene chlo-ride. The combined methylene chloride solutions are dried over magnesium sulfate and concentrated in vacuo to give 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline.

In a similar fashion 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroiso-indoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethylisoindoline, and I-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoincloline are obtained by substitut-ing equivalent amounts of 2-bromomethyl-3,4,5,6-tetrachlorobenzoate, 2-bromo-methyl-3,4,5,6-tetramethylbenzoate, and 2-bromomethyl-3,4.5.6-tetramethoxybenzo-ate, respectively, for 2-bromomethyl-3,4,5,6-tetrafluorobenzoate.

N-Benzyloxycarbonyl-a-methyl-glutamic Acid To a stirred solution of a-methyl-D,L-glutamic acid (10 g, 62 mmol) in 2 N
sodium hydroxide (62 mL) at 0-5 C was added benzyl chloroformate (12.7 g, 74.4 mmol) over 30 min. After the addition was complete the reaction mixture was stirred at room temperature for 3 hours. During this time the pH was maintained at 11 by addition of 2N sodium hydroxide ( 33 mL). The reaction mixture was then extracted with ether (60 mL). The aqueous layer was cooled in an ice bath and then acidified with 4N hydrochloric acid (34 mL) to pH= 1. The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The combined ethyl acetate extracts were washed with brine (60 mL) and dried (MgSO4). The solvent was removed in vacuo to give 15.2 g (83%) of N-benzyloxycarbonyl-a-methylglutamic acid as an oil: I H NMR
(CDC13) S 8.73(m, 5H), 5.77(b, IH), 5.09(s, 2H), 2.45-2.27(m, 4H), 2.0(s, 3H).

In a similar fashion from a-ethyl-D,L-glutamic acid and a-propyl-D,L-glutamic acid, there is obtained N-benzyloxycarbonyl-a-ethylglutamic acid and N-benzyloxy-carbonyl-a-propylglutamic acid, respectively.

N-Benzyloxycarbonyl-a-methyl-glutamic Anhydride A stirred mixture of N-benzyloxycarbonyt-a-methyl-glutamic acid (15 g, 51 mmol) and acetic anhydride (65 mL) was heated at reflux under nitrogen for 30 min.
The reaction mixture was cooled to room temperature and then concentrated in vacuo to afford N-benzylcarbonyl-a-methylglutamic anhydride as an oil (15.7 g) which can be used in next reaction without further purification: 'H NMR (CDC13) S 7.44-7.26 (m, 5H), 5.32-5.30 (m, 2H), 5.11 (s, 1H), 2.69-2.61 (m, 2H), 2.40-2.30 (m, 2H), 1.68 (s, 3H).

In a similar fashion from N-benzyloxycarbonyl-a-ethylglutamic acid and N-benz-yloxycarbonyl-a-propylglutamic acid, there is obtained N-benzylcarbonyl-a-ethyl-glutamic anhydride and N-benzylcarbonyl-a-propylglutamic anhydride, respectively.

N-Benzyloxycarbonyl-a-methylisoglutamine A stirred solution of N-benzylcarbonyl-a-methylglutamic anhydride (14.2 g, 51.5 mmol) in methylene chloride (100 mL) was cooled in an ice bath. Gaseous ammonia was bubbled into the cooled solution for 2 hours. The reaction mixture was stirred at room temperature for 17 hours and then extracted with water (2 x 50 mL). The combined aqueous extracts were cooled in an ice bath and acidified with 4N
hydro-chloric acid (32 mL) to pH 1. The resulting mixture was extracted with ethyl acetate (3 x 80 mL). The combined ethyl acetate extracts were washed with brine (60 mL) and then dried (MgSO4). The solvent was removed in vacuo to give 11.5 g of N-benzyloxycarbonyl-a-amino-a-methylisoglutamine: 'H NMR (CDC13/DMSO) 8 7.35 (m, 5H), 7.01 (s, 1 H), 6.87 (s, 1 H), 6.29 (s, 1 H). 5.04 (s, 2H), 2.24-1.88 (m, 4H), 1.53 (s, 3H).

T. - T

In a similar fashion from N-benzylcarbonyl-a-ethylglutamic anhydride and N-benzylcarbonyl-a-propylglutamie anhydride there is obtained N-benzyloxyearbonyl-a-amino-a-ethylisoglutamine and N-benzyloxycarbonyl-a-amino-a-propylisogluta-mine, respectively.

N-Benzyloxycarbonyl-a-amino-a-methylglutarimide A stirred mixture of N-benzyloxycarbonyl-a-methylisoglutamine (4.60 g, 15.6 mmol), 1,1'-carbonyldiimidazole (2.80 g, 17.1 mmol), and 4-dimethylaminopyridine (0.05 g) in tetrahydrofuran (50 mL) was heated to reflux under nitrogen for 17 hours.
The reaction mixture was then concentrated in vacuo to an oil. The oil was slurried in water (50 mL) for 1 hour. The resulting suspension was filtered and the solid washed with water and air dried to afford 3.8 g of the crude product as a white solid. The crude product was purified by flash chromatography (methylene chloride:ethyl acetate 8:2) to afford 2.3 g (50%) of N-benzyloxycarbonyl-a-amino-a-methylglutarimide as a white solid: mp 150.5-152.5 C; 1H NMR (CDC13) b 8.21 (s, 1H), 7.34 (s, 5H), 5.59 (s, 1H), 5.08 (s, 2H), 2.74-2.57 (m, 3H), 2.28-2.25 (m, 1H), 1.54 (s, 3H); 13C NMR
(CDC13) 5 174.06, 171.56, 154.68, 135.88, 128.06, 127.69, 127.65, 66.15, 54.79, 29.14, 28.70, 21.98; HPLC : Waters Nova-Pak C18 column, 4 micron, 3.9x150 mm, I mL/min, 240nm, 20/80 CH3CN/0.1 % H3P04(aq), 7.56 min (100%); Anal. Calcd For C14H16N204; C, 60.86; H, 5.84; N, 10.14. Found: C, 60.88; H, 5.72; N, 10.07.

In a similar fashion from N-benzyloxycarbonyl-a-amino-a-ethylisoglutamine and N-benzyloxycarbonyl-a-amino-a-propylisoglutamine there is obtained N-benzyloxy-carbonyl-a-amino-a-ethylglutarimide and N-benzyloxycarbonyl-a-amino-a-propyl-glutarimide, respectively.

a-Amino-a-methylglutarimide hydrochloride N-Benzyloxycarbonyl-a-amino-a-methylglutarimide (2.3 g, 8.3 mmol) was dissolved in ethanol (200 mL) with gentle heat and the resulting solution allowed to cool to room temperature. To this solution was added 4N hydrochloric acid (3 mL) followed by 10% Pd/C (0.4 g). The mixture was hydrogenated in a Parr apparatus under 50 psi of hydrogen for 3 hours. To the mixture was added water (50 mL) to dissolve the product. This mixture was filtered through a Celite pad which was washed with water (50 mL). The filtrate was concentrated in vacuo to afford a solid residue. The solid was slurried in ethanol (20 mL) for 30 min. The slurry was filtered to afford 1.38 g (93%) of a-amino-a-methylglutarimide hydrochloride as a white solid: 'H NMR (DMSO-d6) 8 11.25 (s, 1H), 8.92 (s, 3H), 2.84-2.51 (m, 2H), 2.35-2.09 (m, 2H), 1.53 (s, 3H); HPLC, Waters Nova-PakTMC, column, 4 micron, 1 mL/min, 240 nm, 20/80 CH3CN/ 0.1 % H3PO4(aq), 1.03 min (94.6%).

In a similar fashion from N-benzyloxycarbonyl-a-amino-a-ethylglutarimide and N-benzyloxycarbonyl-a-amino-a-propylglutarimide there is obtained a-amino-a-ethylglutarimide hydrochloride and a-amino-a-propylglutarimide hydrochloride, respectively.

3-(3-Nitrophthalimido)-3-methylpiperidine-2,6-dione A stirred mixture of a-amino-a-methylglutarimide hydrochloride (1.2 g, 6.7 mmol), 3-nitrophthalic anhydride (1.3 g, 6.7 mmol), and sodium acetate (0.6 g, 7.4 mmol) in acetic acid (30 mL) was heated to reflux under nitrogen for 6 hours.
The mixture then was cooled and concentrated in vacuo. The resulting solid was slurried in water (30 mL) and methylene chloride (30 mL) for 30 min. The suspension was filtered, the solid was washed with methylene chloride, and dried in vacuo (60 C, <1 mm) to afford 1.44 g (68%) of 3-(3-nitrophthalimido)-3-methylpiperidine-2,6-dione as a off-white solid : mp 265-266.5 C; 1H NMR (DMSO-d6) 8 11.05 (s, IH), 8.31 (dd, J=1.1 and 7.9 Hz, 1H), 8.16-8.03 (m, 2H), 2.67-2.49 (m, 3H), 2.08-2.02 (m, 1H), 1.88 (s, 3H); 13C NMR (DMSO-d6) 8 172.20, 171.71, 165.89, 163.30, 144.19, 136.43, 133.04, 128.49, 126.77, 122.25, 59.22, 28.87, 28.49,-21.04; HPLC, Water Nova-Pa1 C1 column, 4 micron, 1 mL/min, 240nm, 20/80 CH3CN/0.1% H3PO4(aq), 7.38 min(98%). Anal. Calcd For C14H1IN306 : C, 53.00; H, 3.49; N, 13.24. Found : C, 52.77; H, 3.29; N, 13.00.

In a similar fashion from a-amino-a-ethylglutarimide hydrochloride and a-amino-a-propylglutarimide hydrochloride there is obtained 3-(3-nitrophthalimido)-3-ethylpiperidine-2,6-dione and 3-(3-nitrophthalimido)-3-propylpiperidine-2,6-dione, 30. respectively.

3-(3-Aminophthalimido)-3-methylpiperidine-2,6-dione 3-(3-Nitrophthalimido)-3-methylpiperidine-2,6-dione (0.5 g, 1.57 mmol) was dissolved in acetone (250 mL.) with gentle heat and then cooled to room temperature.
To this solution was added 10% Pd/C (0.1 g) under nitrogen. The mixture was hydrogenated in a Parr apparatus at 50 psi of hydrogen for 4 hours. The mixture then was filtered through Celite and the pad washed with acetone (50 mL). The filtrate was concentrated in vacuo to yield a yellow solid. The solid was slurried in ethyl acetate (10 mL) for 30 minutes. The slurry then was filtered and dried (60 C, <1 mm) to afford 0.37 g (82%) of 3-(3-aminophthalimido)-3-methylpiperidine-2,6-dione as a yellow solid: mp 268-269 C; 1H NMR (DMSO-d6) 8 10.98 (s, 1H), 7.44 (dd, J=7.1 and 7.3 Hz, 1 H), 6.99 (d, J=8.4 Hz, I H), 6.94 (d, J=6.9 Hz, 1 H), 6.52 (s, 2H), 2.71-2.47 (m, 3H), 2.08-1.99 (m, 1H), 1.87 (s, 3H); 13C NMR (DMSO-d6) S 172.48, 172.18, 169.51, 168.06, 146.55, 135.38, 131.80, 121.51, 110.56, 108.30, 58.29, 29.25, 28.63, 21.00; HPLC, Water Nova-Pak/C18 column, 4 micron, 1 mL/min, 240 rim, 20/80 CH3CN/0.1%H3PO4(aq), 5.62 min (99.18%). Anal. Calcd For C14H13N304 : C, 58.53; H, 4.56; N, 14.63. Found : C, 58.60; H, 4.41; N, 14.36.

In a similar fashion from 3-(3-nitrophthalimido)-3-ethylpiperidine-2,6-dione and 3-(3-nitrophthal imido)-3-propylpiperidine-2,6-di one there is obtained 3-(3-aminophthal-imido)-3-ethylpiperidine-2,6-dione and 3-(3-aminophthalimido)-3-propylpiperidine-2,6-dione, respectively.

Methyl 2-bromomethyl-3-nitrobenzoate A stirred mixture of methyl 2-methyl-3-nitrobenzoate(17.6 g, 87.1 mmol) and N-bromosuccinimide (18.9 g, 105 mmol) in carbon tetrachloride (243 mL) was heated under gentle reflux with a 100 W light bulb situated 2 cm away shining on the reac-tion mixture overnight. After 18 hours, the reaction mixture was cooled to room temperature and filtered. The filtrate was washed with water (2 x 120 mL), brine(120 mL), and dried (MgSO4). The solvent was removed in vacuo to give a yellow solid.
The product was purified by flash chromatography (hexane:ethyl acetate 8:2) to give 22 g (93%) of methyl 2-bromomethyl-3-nitrobenzoate as a yellow solid: mp 69-72 C;
1H NMR (CDCl3) S 8.13-8.09 (dd, J=1.36 and 7.86 Hz, 1H), 7.98-7.93 (dd, J=1.32 and 8.13 Hz, 1H), 7.57-7.51 (t, J=7.97Hz, IH), 5.16 (s, 2I1), 4.0 (s, 3H); 13C
NMR

(CDC13) 6 65.84, 150.56, 134.68, 132.64, 132.36, 129.09, 53.05, 22.70; HPLC
Waters Nova-Pak C, 8 column, 4 micron, 1 mL/min, 240nm, 40/60 CH3CN/0.1%H3PO4(aq), 8.2 min 99 %. Anal. Caled for CgHgNO4Br: C, 39.44; H, 2.94; N, 5.11, Br, 29.15. Found: C, 39.51; H, 2.79; N, 5.02; Br, 29.32.

3-(1-Oxo-4-nitroisoindolin-1-yl)-3-mcthylpiperidine-2,6-dione To a stirred mixture of a-amino-a-methylglutarimide hydrochloride (2.5g, 14.0 mmol) and methyl 2-bromomethyl-3-nitrobenzoate(3.87g, 14.0 mmol in dimethyl-formamide (40 mL) was added triethylamine (3.14g, 30.8 mmol). The resulting 10 mixture was heated to reflux under nitrogen for 6 hours. The mixture was cooled and then concentrated in vacuo. The resulting solid was slurried in water (50 mL) and CH2Cl2 for 30 min. The slurry was filtered, the solid washed with methylene chloride, and dried in vacuo (60 C , <lmm) to afford 2.68 g (63%) of 3-(1-oxo-4-nitroisoindo-lin-1-yl)-3-methylpiperidine-2,6-dione as a off-white solid : mp 233-235 C;
'H NMR
15 (DMSO-d6) S 10.95 (s, 1H), 8.49-8.46 (d, J=8.15 Hz, 111), 8.13-8.09 (d, J=7.43 Hz, 1H), 7.86-7.79 (t, J= 7.83 Hz, 1H), 5.22-5.0 (dd, J=19.35 and 34.6 Hz, 2H), 2.77-2.49 (m, 3H), 2.0-1.94 (m, 1H), 1.74 (S, 3H); 13C NMR (DMSO-d6) S 173.07, 172.27, 164.95, 143.15, 137.36, 135.19, 130.11, 129.32, 126.93, 57.57, 48.69, 28.9, 27.66, 20.6; HPLC, Waters Nova-Pak C,8 column, 4 micron, 1 mL/min, 240nm, 20/80 20 CH3CN/0.1%H3P04(aq), 4.54 min 99.6%. Anal. Calcd for C14H13N305: C, 55.45;
H, 4.32; N, 13.86. Found: C, 52.16; H, 4.59; N. 12.47.

By substituting equivalent amounts of a-amino-a-ethylglutarimide hydrochloride and a-amino-ct-propylglutarimide hydrochloride for a-amino-a-methylglutarimide hydrochloride, there is obtained respectively 3-(1-oxo-4-nitroisoindolin-1-yl)-ethylpiperidine-2,6-dione and 3-(1-oxo-4-nitroisoindolin-1-yl)-3-propylpiperidine-2,6-dione.

3-(1-Oxo-4-aminoisoindolin-1-yl)-3-methylpiperidine-2,6-dione 3-(1-Oxo-4-nitroisoindolin-1-yl)-3-methylpiperidine-2,6-dione (1.0 g, 3.3 mmol) was dissolved in methanol (500 mL) with gentle heat and allowed to cool to room temperature. To this solution was added 10% Pd/C (0.3 g) under nitrogen. The mixture was hydrogenated in a Parr apparatus at 50 psi of hydrogen for 4 hours. The T i mixture was filtered through Celite and the Celite washed with methanol (50 mL).
The filtrate was concentrated in vacuo to an off white solid. The solid was slurried in methylene chloride (20 mL) for 30 min. The slurry was then filtered and the solid dried (60 C, <1 mm) to afford 0.54 g (60%) of 3-(1-oxo-4-aminoisoindolin-1-yl)-methylpiperidine-2,6-dione as a white solid: mp 268-270 C; 'H NMR (DMSO-d6) S
10.85 (s, 111), 7.19-7.13 (t, J=7.63 Hz, I H), 6.83-6.76 (m, 2H), 5.44 (s, 2H), 4.41(s, 2H), 2.71-2.49 (m, 3H), 1.9-1.8 (m, 111), 1.67 (s, 3H); 13C NMR (DMSO-d6) 8 173.7, 172.49, 168.0, 143.5, 132.88, 128.78, 125.62, 116.12, 109.92, 56.98, 46.22, 29.04, 27.77, 20.82; HPLC, Waters Nova-Pak/C18 column, 4 micron, 1 mL/min, 240 nm, 20/80 CH3CN/0.l%H3P04(aq), 1.5 min (99.6%); Anal. Calcd for C14H15N303 : C, 61.53; H, 5.53; N, 15.38. Found : C, 58.99; H, 5.48; N, 14.29.

From 3-(1-oxo-4-nitroisoindolin-1-yl)-3-ethylpiperidine-2,6-dione and 3-(1-oxo-nitroisoindolin-1-yl)-3-propylpiperidine-2,6-dione there is similarly obtained 3-(]-oxo-4-aminoisoindolin-1-yl)-3-ethylpiperidine-2,6-dione and 3-(1-oxo-4-aminoiso-indolin- l -yl)-3-propylpiperidine-2,6-dione, respectively.

S-4-Amino-2-(2,6-dioxopiperid-3-yl)isoindoline-1,3-dione A. 4-Nitro-N-et hoxycarbonylphthal imide Ethyl chloroformate (1.89 g, 19.7 mmol) was added dropwise over 10 min to a stirred solution of 3-nitrophthalimide (3.0 g, 15.6 mmol) and triethylamine (1.78 g.
17.6 mmol) in dimethylformamide (20 mL) at 0-5 C under nitrogen. The reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The mixture was then slowly added to an agitated mixture of ice and water (60 mL).
The resulting slurry was filtered and the solid was crystallized from chloroform (15 mL) and pet ether (15 mL) to afford 3.1 g (75%) of the product as an off-white solid: mp 100-100.5 C; 'H NMR (CDC13) 8 8.25(d, J=7.5 Hz, 1H), 8.20(d, J=8.0 Hz, 1H), 8.03(t, J=7.9 Hz, 1H), 4.49(q, J=7.1 Hz, 2H), 1.44(t, J=7.2 Hz, 3H); 13C NMR
(CDC13) 8 161.45, 158.40, 147.52, 145.65, 136.60, 132.93, 129.65, 128.01, 122.54;
64.64, 13.92; HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, I mL/min, 240 nm, 30/70 CH3CN/0.1%H3P04(aq), 5.17 min(98.11%); Anal. Calcd for C1,H8N206:
C, 50.00; H, 3.05; N, 10.60. Found : C, 50.13; H, 2.96; N, 10.54.

B. i-Butyl N-(4-nitrophthaloyl)-L-glutamine A stirred mixture of 4-nitro-N-ethoxycarbonylphthalimide (1.0 g, 3.8 mmol), L-glutamine t-butyl ester hydrochloride (0.90 g, 3.8 rnmol) and triethylamine (0.54 g, 5.3 mmol) in tetrahydrofuran (30 mL) was heated to reflux for 24 hours. The 5 tetrahydrofuran was removed in vacuo and the residue was dissolved in methylene chloride (50 mL). The methylene chloride solution was washed with water (2x15 mL), brine (15 mL) and then dried (sodium sulfate). The solvent was removed in vacuo and the residue was purified by flash chromatograph (7:3 methylene chloride:ethyl acetate) to give 0.9 g (63%) of a glassy material: IH NMR
(CDC13) 6 8.15(d, J=7.9 Hz, 2H), 7.94(t, J=7.8 Hz, 1H), 5.57(b, 2H), 4.84(dd, J=5.1 and 9.7 Hz, 1H), 2.53-2.30(m, 4H), 1.43(s, 9H); HPLC, Wasters Nova-Pal7C18, 3.9x150 mm, 4 micron, I mL/min, 240 run, 30/70 CH3CN/0.1 %H3PO4(aq), 6.48 min(99.68%);
Chiral Analysis, Daicel Chiral Pak"AD, 0.4x25 Cm, I mL/min, 240 nm, 5.32 min(99.39%);
Anal. Calcd for C17H19N3O7 : C, 54.11; H, 5.08; N, 11.14. Found : C, 54.21; H, 5.08;
N, 10.85.

C. N-(4-Nitrophthaloyl)-L-glutamine Hydrogen chloride gas was bubbled into a stirred 5 C solution of t-butyl N-(4-nitrophthaloyl)-L-glutamine (5.7 g, 15.1 mmol) in methylene chloride (100 mL) for min. The mixture was then stirred at room temperature for 16 hours. Ether (50 20 mL) was added and the resulting mixture was stirred for 30 min. The resulting slurry was filtered to yield 4.5 g of crude product as a solid, which was used directly in the next reaction : 1 H NMR (DMSO-d6) 5 8.36(dd, J=0.8 and 8.0 Hz, 1 H), 8.24(dd, J=0.8 and 7.5 Hz, 1 H), 8.11(t, J=7.9 Hz, 1 H), 7.19(b, I H), 6-72(b, 1 H), 4.80(dd, J=3.5 d an 8.8 Hz, 1H), 2.30-2.10(m, 4H).

25 D. (S)-2-(2,6-dioxo(3 piperidyl))-4-nitroisoindoline-1,3-dione A stirred suspension of N-(4-nitrophthaloyl)-L-glutamine (4.3 g, 13.4 mmol) in anhydrous methylene chloride (170 mL) was cooled to -40 C (IPA/dry ice bath).
Thionyl chloride (1.03 mL, 14.5 mmol) was added dropwise to the mixture followed by pyridine (1.17 mL, 14.5 mmol). After 30 minutes. triethylamine (2.06 mL, 14.8 mmol) was added and the mixture was stirred at -30 to -40 C for 3 hours. The mixture was allowed to warm to room temperature, filtered and washed with methylene chloride to afford 2.3 g (57%) of the crude product.
Recrystallization from acetone (300 mL) afforded 2 g of the product as a white solid : mp 259.0-284.0 C(dec.); 'H NMR (DMSO-d6) 6 11.19(s, 1H), 8.34(d, J=7.8 Hz, 1H), 8.23(d, J=7.1 Hz, 1H), 8.12(t, J=7.8 Hz, 1H), 5.25-5.17(dd, J=5.2 and 12.7 Hz, 1H), 2.97-2.82(m, 1H), 2.64-2.44(m, 2H), 2.08-2.05(m, 1H); '3C NMR (DMSO-d6) S 172.67, 169.46, 165.15, 162.50, 144.42, 136.78, 132.99, 128.84, 127.27, 122.53, 49.41, 30.84, 21.71; HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, 1mL/min, 240 rim, 10/90 CH3CN/0.1%H3P04(aq) 4.27 min(99.63%); Anal. Calcd for C13H9N306 : C, 51.49; H, 2.99; N, 13.86. Found : C, 51.67; H, 2.93; N, 13.57.

E. S-4-Amino-2-(2,6-dioxopiperid-3 yl)isoindoline-l,3-dione.
A mixture of (S)-3-(4'-nitrophthalimido)-piperidine-2,6-dione (0.76 g, 2.5 mmol) and 10%Pd/C (0.3 g) in acetone (200 mL) was hydrogenated in a Parr-Shaker apparatus at 50 psi of hydrogen for 24 hours. The mixture was filtered through celite and the filtrate was concentrated in vacuo. The solid residue was slurried in hot ethyl acetate for 30 min and filtered to yield 0.47 g (69%) of the product as a yellow solid :
mp 309-310 C; 'H NMR (DMSO-d6) 6 11.10 (s, 1H), 7.47(dd, J=7.2 and 8.3 Hz, 1H), 7.04-6.99(dd, J=6.9 and 8.3 Hz, 2H), 6.53(s, 2H), 5.09-5.02(dd, J=5.3 and 12.4 Hz, 1H), 2.96-2.82(m, 1H), 2.62-2.46(m, 2H), 2.09-1.99(m, 1H); 13C NMR (DMSO-d6) 6 172.80, 170.10, 168.57, 167.36, 146.71, 135.44, 131.98, 121.69, 110.98, 108.54, 48.48, 30.97, 22.15; HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, 1 mL/min, 240 rim, 15/85 CH3CN/0.1 %H3P04(aq) 4.99 min(98.77%); Chiral analysis, Daicel Chiral Pak AD, 0.46x25 cm, 1 mL/min, 240 rim, 30/70 Hexane/IPA 9.55 min (1.32%), 12.55 min (97.66%); Anal. Calcd for C13H11N304 : C, 57.14; H, 4.06;
N, 15.38. Found : C, 57.15; H, 4.15; N, 14.99.

R-4-Amino-2-(2,6-dioxopiperid-3-yl))isoindoline-1,3-dione A. t-Butyl N-(4-nitrophthaloyl)-D-glutamine A stirred mixture of 4-nitro-N-ethoxycarbonyl-phthalimide (5.9 g, 22.3 mmol), D-glutamine t-butyl ester (4.5 g, 22.3 mmol) and triethylamine (0.9 g, 8.9 mmol) in tetrahydrofuran (100 mL) was refluxed for 24 hours. The mixture was diluted with methylene chloride (100 mL) and washed with water (2 x 50 mL), brine (50 mL) and then dried. The solvent was removed in vacuo and the residue was purified by flash chromatography (2% CH3OH in methylene chloride) to afford 6.26 g (75%) of the product as a glassy material : 'H NMR (CDCl3) 8 8.12(d, J=7.5 Hz, 2H), 7.94(dd, J=7.9 and 9.1 Hz, IH), 5.50(b, 1H), 5.41(b, 1H), 4.85(dd, J=5.1 and 9.8 Hz, IH), 2.61-2.50(m, 2H), 2.35-2.27(m,2H), 1.44(s, 9H); 13C NMR (CDC13) S 173.77, 167.06, 165.25, 162.51, 145.07, 135.56, 133.78, 128.72, 127.27, 123.45, 83.23, 53.18, 32.27, 27.79, 24.42,; HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, 1 mL/min, 240 nm, 25/75 CH3CN/0.1 %H3P04(aq) 4.32 min(99.74%); Chiral analysis, Daicel Chiral Pak AD, 0.46x25 cm, 1 mL/min, 240 rim, 55/45 Hexane/IPA 5.88 min(99.68%); Anal. Caled for C17H19N307 : C, 54.11; H, 5.08; N, 11.14. Found :
C, 54.25; H, 5.12; N, 10.85.

B. N-(4-Nitrophthaloyl)-D-glutamine Hydrogen chloride gas was bubbled into a stirred 5 C solution of t-butyl N-(4-nitrophthaloyl)-D-glutamine (5.9 g, 15.6 mmol) in methylene chloride (100 mL) for 1 hour then stirred at room temperature for another hour. Ether (100 mL) was added and stirred for another 30 minutes. The mixture was filtered, the solid was washed with ether (60 mL) and dried (40 C, <lmm Hg) to afford 4.7 g (94%) of the product:
1H NMR (DMSO-d6) 6 8.33(d, J=7.8 Hz, 111), 8.22(d, J=7.2 Hz, IH), 8.11(t, J=7.8 Hz, 1 H), 7.19(b, I H), 6.72(b, 1 H), 4.81(dd, J=4.6 and 9.7 Hz, 1 H), 2.3 9-2.12(m, 4H);
13C NMR (DMSO-d6) 6 173.21, 169.99, 165.41, 162.73, 144.45, 136.68, 132.98, 128.80, 127.23, 122.52, 51.87, 31.31, 23.87.

C. (R)-2-(2,6-dioxo(3 piperidyl))-4-nitroisoindoline-1,3-dione.
A stirred suspension of N-(4'-nitrophthaloyl)-D-glutamine (4.3 g, 13.4 mmol) in anhydrous methylene chloride (170 mL) was cooled to -40 C with isopropanol/dry ice bath. Thionyl chloride (1.7 g, 14.5 mmol) was added dropwise followed by pyridine (1.2 g, 14.5 mmol). After 30 min, triethylamine (1.5 g, 14.8 mmol) was added and the mixture was stirred at -30 to -40 C for 3 hours. The mixture was filtered, the solid washed with methylene chloride (50 mL) and dried (60 C, <lmm Hg) to give 2.93 g of the product. Another 0.6 g of the product was obtained from the methylene chloride filtrate. Both fractions were combined (3.53 g) and recrystallized from acetone (450 mL) to afford 2.89 g (71 %) of the product as a white solid: mp 256.5-257.5 C; 1H NMR (DMSO-d6) 8 11.18(s, 1H), 8.34(dd, J=0.8 and 7.9 Hz, IH), 8.23(dd, J=0.8 and 7.5 Hz, 1H), 8.12(t, J=7.8 Hz, IH), 5.22(dd, J=5.3 and 12.8 Hz, IH), 2.97-2.82(m, 1H), 2.64-2.47(m, 2H), 2.13-2.04(m, IH); 13C NMR
(DMSO-d6) 6 172.66, 169.44, 165.14, 162.48, 144.41, 136.76, 132.98, 128.83, 127.25, 122.52, 49.41, 30.83, 21.70; HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 r - t micron, 1 mL/min, 240 nm, 10/90 CH3CN/0.1%H3PO4(aq) 3.35 min(100%); Anal.
Calcd for C13H9N306 : C, 51.49; H, 2.99; N, 13.86. Found : C, 51.55; H, 2.82;
N, 13.48.

D. (R)-4-Amino-2-(2,6-dioxopiperid-3 yl)isoindoline-J,3-dione A mixture of R-3-(4'-nitrophthalimido)-piperidine-2,6-dione (1.0 g, 3.3 mmol) and 10% Pd/C (0.2 g) in acetone (250 mL) was hydrogenated in a Parr-Shaker apparatus at 50 psi of hydrogen for 4 hours. The mixture was filtered through celite and the fitrate was concentrated in vacuo. The resulting yellow solid was slurried in hot ethyl acetate (20 mL) for 30 min to give after filtration and drying 0.53 g (59%) of the product as a yellow solid : mp 307.5-309.5 C; 'H NMR (DMSO-d6) S 11.06(s, 1H), 7.47(dd, J=7.0 and 8.4 Hz, 1H), 7.02(dd, J=4.6 and 8.4 Hz, 2H), 6.53(s, 2H), 5.07(dd, J=5.4 and 12.5 Hz, 1H), 2.95-2.84(m, IH), 2.62-2.46(m, 2H), 2.09-1.99(m, IH); J3C NMR (DMSO-d6) S 172.78, 170.08, 168.56, 167.35, 146.70, 135.43, 131.98, 121.68, 110.95, 108.53, 48.47, 30.96, 22.14; HPLC, Waters Nove-Pak/C18, 3.9x150 mm, 4 micron, 1 mL/min, 240 nm, 10/90 CH3CN/0.1 %H3PO4(aq) 3.67 min(99.68%);
Chiral analysis, Daicel Chiral Pak AD, 0.46x25 cm, I mL/min, 240 nm, 30/70 Hexane/ IPA 7.88min (97.48%); Anal. Calcd for C13H,IN304 : C, 57.14; H, 4.06;
N, 15.38. Found : C, 57.34; H, 3.91; N, 15.14.

3-(4-Amino-l-oxoisoindolin-2-yl)piperidine-2,6-dione A. Methyl 2-bromomethyl-3-nitrobenzoate A stirred mixture of methyl 2-methyl-3-nitrobenzoate (14.0 g, 71.7 mmol) and N-bromosuccinimide (15.3 g, 86.1 mmol) in carbon tetrachloride (200 mL) was heated under gentle reflux for 15 hours while a 100W bulb situated 2 cm away was shining on the flask. The mixture was filtered and the solid was washed with methylene chloride (50 mL). The filtrate was washed with water (2x100 mL), brine (100 mL) and dried. The solvent was removed in vacuo and the residue was purified by flash chromatography (hexane/ethyl acetate, 8/2) to afford 19 g (96%) of the product as a yellow solid : mp 70.0-71.5 C; 'H NMR (CDCI3) S 8.12-8.09(dd, J=1.3 and 7.8 1-Iz, 1H), 7.97-7.94(dd, J=1.3 and 8.2 Hz, 1H), 7.54(t, J=8.0 Hz, IH), 5.15(s, 21-I), 4.00(s, 3H); 13C NMR (CDC13) S 165.85, 150.58, 134.68, 132.38, 129.08, 127.80, 53.06, 22.69; HPLC, Water Nove-Pak/C18, 3.9x150 mm, 4 micron, 1 mL/min, 240 nm, 40/60 CH3CN/0.1 %H3PO4(aq) 7.27 min(98.92%); Anal. Calcd for CgHgNO4Br : C, 39.44; H, 2.94; N, 5.11; Br, 29.15. Found : C, 39.46; H, 3.00;
N, 5.00; Br, 29.11.

B. t-Butyl N-(1-oxo-4-nitroisoindolin-2 yl)-L-glutamine Triethylamine (2.9 g, 28.6 mmol) was added dropwise to a stirred mixture of methyl 2-bromomethyl-3-nitrobenzoate (3.5 g, 13.0 mmol) and L-glutamine t-butyl ester hydrochloride (3.1 g, 13.0 mmol) in tetrahydrofuran (90 mL). The mixture was heated to reflux for 24 hours. To the cooled mixture was added methylene chloride (150 mL) and the mixture was washed with water (2 x 40 mL), brine (40 mL) and dried. The solvent was removed in vacuo and the residue was purified by flash chromatography (3% CH3OH in methylene chloride) to afford 2.84 g (60%) of crude product which was used directly in the next reaction: 'H NMR (CDC13) 8 8.40(d, J=8.1 Hz, I H), 8.15(d, J=7.5 Hz, 1H), 7.71(t, J=7.8 Hz, III), 5.83(s, I H), 5.61(s, 1H), 5.12(d, J=19.4 Hz, 1H), 5.04-4.98(m,1H), 4.92(d, J=19.4 Hz, 1H), 2.49-2.22(m, 4H), 1.46(s, 9H); HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, I mL/min, 240 nm, 25/75 CH3CN/0.1%H3P04(aq) 6.75 min(99.94%).

C. N-(1-Oxo-4-nitroisoindolin-2 yl)-L-glutamine Hydrogen chloride gas was bubbled into a stirred 5 C solution of t-butyl N-(1-oxo-4-nitro-isoindolin-2-yl)-L-glutamine (3.6 g, 9.9 mmol) in methylene chloride (60 mL) for 1 hour. The mixture was then stirred at room temperature for another hour.
Ether (40 mL) was added and the resulting mixture was stirred for 30 minutes.
The slurry was filtered, washed with ether and dried to afford 3.3 g of the product : 'H
NMR (DMSO-d6) S 8.45(d, J=8.1 Hz, I H), 8.15(d, J=7.5 Hz, 1H), 7.83(t, J=7.9 Hz, IH), 7.24(s, IH), 6.76(s, I H), 4.93(s, 2H), 4.84-4.78(dd, J=4.8amd 10.4 Hz, 1H), 2.34-2.10(m, 4H); 13C NMR (DMSO-d6) 6 173.03, 171.88, 165.96, 143.35, 137.49, 134.77, 130.10, 129.61, 126.95, 53.65, 48.13, 31.50, 24.69; Anal. Calcd for C13H13N306 : C, 50.82; H, 4.26; N, 13.68. Found : C, 50.53; H, 4.37; N, 13.22.

D. (S)-3-(1 -Oxo-4-nitroisoindolin-2 yl)piperidine-2, 6-dione A stirred suspension mixture of N-(I-oxo-4-nitroisoindolin-2-yl)-L-glutamine (3.2 g, 10.5 mmol) in anhydrous methylene chloride (150 mL) was cooled to -40 C
with isopropanol/dry ice bath. Thionyl chloride (0.82 mL, 11.3 mmol) was added dropwise to the cooled mixture followed by pyridine (0.9 g, 11.3 mmol). After T T

min, triethylamine (1.2 g, 11.5 mmol) was added and the mixture was stirred at -30 to -40 C for 3 hours. The mixture was poured into ice water (200 mL) and the aqueous layer was extracted with methylene chloride (40 mL). The methylene chloride solution was washed with water (2 x 60 mL), brine (60 mL) and dried. The solvent was removed in vacuo and the solid residue was slurried with ethyl acetate (20 mL) to give 2.2 g (75%) of the product as a white solid: mp 285 C; 'H NMR (DMSO-d6) S
11.04(s, 1H), 8.49-8.45(dd, J=0.8 and 8.2 Hz, IH), 8.21-8.17(dd, J=7.3 Hz, IH), 7.84(t, J=7.6 Hz, 1H), 5.23-5.15(dd, J=4.9 and 13.0 Hz, 1H), 4.96(dd, J=19.3 and 32.4 Hz, 2H), 3.00-2.85(m, 1H), 2.64-2.49(m, 2H), 2.08-1.98(m, 1H); 13C NMR (DMSO-d6) 8 172.79, 170.69, 165.93, 143.33, 137.40, 134.68, 130.15, 129.60, 127.02, 51.82, 48.43, 31.16, 22.23; HPLC, Waters Nove-Pak/C 18, 3.9x 150 mm, 4 micron, I
mL/min, 240 rim, 20/80 CH3CN/0.1 %H3P04(aq) 3.67 min(100%); Anal. Calcd for Cf3H1 IN305 : C, 53.98; H, 3.83; N, 14.53. Found : C, 53.92; H, 3.70; N, 14.10.

E. (S)-3-(] -Oxo-4-aminoisoindolin-2 yl)piperidine-2, 6 dione A mixture of (S)-3-(1-oxo-4-nitroisoindolin-2-yl)piperidine-2,6-dione (1.0 g, 3.5 mmol) and 10% Pd/c (0.3 g) in methanol (600 mL) was hydrogenated in a Parr-Shaker apparatus at 50 psi of hydrogen for 5 hours. The mixture was filtered through Celite and the filtrate was concentrated in vacuo. The solid was slurried in hot ethyl acetate for 30 min , filtered and dried to afford 0.46 g (51%) of the product as a white solid: mp 235.5-239 C; 'H NMR (DMSO-d6) S 11.01(s, 1H), 7.19(t, J=7.6 Hz, IH), 6.90(d, J=7.3 Hz, IH), 6.78(d, J=7.8 Hz, IH), 5.42(s, 2H), 5.12(dd, J=5.1 and 13.1 Hz, 1H), 4.17(dd, J=17.0 and 28.8 Hz, 2H), 2.92-2.85(m, 1H). 2.64-2.49(m, IH), 2.34-2.27(m, 1H), 2.06-1.99(m, IH); 13C NMR (DMSO-d6) 6 172.85, 171.19, 168.84, 143.58, 132.22, 128.79, 125.56, 116.37, 110.39, 51.48, 45.49, 31.20, 22.74;
HPLC, Waters Nova-Pak/C18, 3.9x150 mm, 4 micron, 1 mL/min, 240 rim, 10/90 CH3CN/0.1%H3P04(aq) 0.96 min(100%); Chiral analysis, Daicel Chiral Pak AD, 40/60 Hexane/IPA, 6.60 min(99.42%); Anal. Calcd for C13H13N303 : C, 60.23; H, 5.05; N. 16.21. Found : C, 59.96; H, 4.98; N, 15.84.

3-(4-Amino-l -oxoisoindolin-2y1)-3-methylpiperidine-2,6-dione A. N-Benzyloxycarbonyl-3-amino-3-methylpiperidine-2, 6-dione A stirred mixture of N-benzyloxycarbonyl-a-methyl-isoglutamine (11.3 g, 38.5 mmol), 1,1'-carbonyldiimidazole (6.84 g, 42.2 mmol) and 4-dimethylaminopyridine (0.05 g) in tetrahydrofuran (125 mL) was heated to reflux under nitrogen for 19 hours. The reaction mixture was concentrated in vacuo to an oil. The oil was slurried in water (50 mL) for l hour then filtered, washed with water, air dried to afford 7.15 g of white solid. The crude product was purified by flash chromatography (2:8 ethyl acetate:methylene chloride) to afford 6.7 g (63%) of the product as a white solid : mp 151-152 C; 'H NMR (CDCl3) 8 8.24 (s, IH), 7.35 (s, 5H), 5.6 (s, 1H), 5.09 (s, 2H), 2.82-2.53 (m, 3H), 2.33-2.26 (m, I1-I), 1.56 (s, 3H); 13C
NMR (CDC13) 6 174.4, 172.4, 154.8, 136.9, 128.3, 127.8, 127.7, 65.3, 54.6, 29.2, 29.0, 22.18; HPLC : Waters Nova-Pak/C18 column, 4 micron, 3.9x150 mm, lml/min, 240nm, 20/80 CH3CN/H3PO4(aq), 6.6 min, 100%). Anal. Calcd for C14H16N204.
Theory: C, 60.86; H, 5.84; N, 10.14. Found: C, 60.94; H, 5.76; N, 10.10.

B. 3-Amino-3-methylpiperidine-2, 6-dione.
N-benzyloxycarbonyl-3-amino-3-methylpiperidine-2, 6-dione (3.0 g, 10.9 mmol) was dissolved in ethanol (270 mL) with gentle heat and then cooled to room temperature. To this solution was added 4 N HCI (7 mL) followed by 10% Pd/C
(0.52 g). The mixture was hydrogenated under 50 psi of hydrogen for 3 hours.
To the mixture was then added water (65 mL) to dissolve the product. The mixture was filtered through a celite pad and the celite pad washed with water (100 mL).
The filtrate was concentrated in vacuo to a solid residue. This solid was slurried in ethanol (50 mL) for 30 min. The slurry was filtered to afford 3.65 g (94%) of the product as a white solid: 'H NMR (DMSO-d6) 6 11.25 (s, 1H), 8.9 (s, 3H), 2.87-2.57 (m, 2H), 2.35-2.08 (in, 2H), 1.54 (s, 3H); HPLC (Waters Nova-Pak/C,8 column, 4 micron, I
ml/min, 240 nm, 15/85 CH3CN/ H3P04(aq), 1.07 min, 100%).

C. 3-Methyl-3-(4-nitro-l-oxoisoindolin-2 yl)piperidine-2, 6-dione To a stirred mixture of a-amino-a-methyl-glutarimide hydrochloride (2.5 g, 14.0 mmol) and methyl 2-bromomethyl-3-nitro benzoate (3.87 g, 14 mmol in dimethylformamide (40 mL) was added triethylamine (3.14 g, 30.8 mmol) under nitrogen. The mixture was heated to reflux for 6 hours. The mixture was cooled and r , T

then concentrated in vacuo. The solid residue was slurried in water (50 rnL) and methylene chloride for 30min. The slurry was filtered and the solid washed with methylene chloride and dried (60 C , <lmm). Recrystallization from methanol (80 mL) yielded 0.63g (15%) of the product as an off white solid: mp 195-197 C; 1H
NMR (DMSO-d6) 6 10.95 (s, 111), 8.49-8.46 (d, J = 8.2 Hz, 1H), 8.13-8.09 (d, J
= 7.4 Hz, IH), 7.86-7.79 (t, J = 7.8 Hz, IH), 5.22-5.0 (dd, J = 19.4 and 34.6 Hz, 2H), 2.77-2.49 (m, 3H), 2.0-1.94 (m, IH), 1.74 (S, 3H); 13C NMR (DMSO-d6) S 173.1, 172.3, 165.0, 143.2, 137.4, 135.2, 130.1, 129.3, 126.9, 57.6, 48.7, 28.9, 27.7, 20.6;
HPLC
(Waters Nova-Pak/C18 column, 4micron, I ml/min, 240nm, 20/80 CH3CN/H3PO4(8q), 4.54 min, 99.6%); Anal Calcd. For C14H13N305; C, 55.45; H, 4.32; N, 13.86.
Found:
C, 55.30; H, 4.48; N, 13.54.

D. 3-Methyl-3-(4-amino-l -oxoisoindolin-2yl)piperidin a-2,6-dione.
3-Methyl-3-(4-nitro-l-oxoisoindolin-2-yl)piperidine-2,6-dione (1.0 g, 3.3 mmol) was dissolved in methanol (500 mL) with gentle heat and then cooled to room temperature. To this solution was added 10% Pd/C (0.3 g) under nitrogen. The mixture was hydrogenated in a Parr-Shaker apparatus at 50 psi of hydrogen for hours. The mixture was filtered through celite pad and the celite pad washed with methanol (50 mL). The filtrate was concentrated in vacuo to a off white solid.
The solid was slurried in methylene chloride (20 mL) for 30 min. The slurry was filtered and the solid dried (60 C, <1 mm). The solid was to recrystallized from methanol (3 times, 100 mL/time) to yield 0.12 g (13.3%) of the product as a white solid:
mp 289-292 C; 1H NMR (DMSO-d6) S 10.85 (s, 1H), 7.19-7.13 (t, J = 7.6 Hz, 1H), 6.83-6.76 (m, 2H), 5.44 (s, 2H), 4.41(s, 2H), 2.71-2.49 (m, 3H), 1.9-1.8 (m, 1H), 1.67 (s, 3H);
13C NMR (DMSO-d6) S 173.7, 172.5, 168.0, 143.5, 132.9, 128.8, 125.6, 116.1, 109.9, 57.0, 46.2, 29.0, 27.8, 20.8; HPLC (Waters Nova-Pak/C18 column, 4 micron, I
ml/min, 240 nm, 20/80 CH3CN/H3P04(aq), 1.5 min, 99.6%); Anal. Calcd, For C14H15N303; C, 61.53; H, 5.53; N, 15.38. Found: C, 61.22; H, 5.63; N, 15.25.

Tablets, each containing 50 mg of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, can be prepared in the following manner:

Constituent s (for 1000 tablets) 1,3-dioxo-2-(2,6-dioxo-piperidin-3-yl)-5-amino-isoindoline .....................................50.0 g lactose ............................................50.7 g wheat starch .................................... 7.5 g polyethylene glycol 6000 ................ 5.0 g talc ................................................... 5.0 g magnesium stearate ......................... 1.8 g demineralized water ........................ q.s.

The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
The active ingredient, lactose, talc, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspen-sion is added to a boiling solution of the polyethylene glycol in 100 mL of water. The resulting paste is added to the pulverulent substances and the mixture is granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets, each containing 100 mg of 1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, can be prepared in the following manner:

Constituents (for 1000 tablets) 1,3-dioxo-2-(2,6-dioxo-piperidin-3-yl)-5-amino-isoindoline ................................... 100.0 g lactose ..........................................100.0 g wheat starch .................................. 47.0 g magnesium stearate ........................ 3.0 g All the solid ingredients are first forced through a sieve of 0.6 mm mesh width. The active ingredient, lactose, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspension is added to 100 mL of boiling water. The resulting paste is added to the pulverulent substances and the mixture is granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets for chewing, each containing 75 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, can be prepared in the following manner:

Composition (for 1000 tablets) 1-oxo-2-(2,6-dioxo-piperidin-3 -y 1)-4-amino-isoindoline .................................... 75.0 g mannitol .......................................230.0 g lactose ..........................................150.0 g talc ................................................ 21.0 g glycine ........................................... 12.5 g stearic acid .................................... 10.0 g saccharin ........................................ 1.5 g 5% gelatin solution ....................... q.s.

All the solid ingredients are first forced through a sieve of 0.25 mm mesh width. The mannitol and the lactose are mixed, granulated with the addition of gelatin solution, forced through a sieve of 2 mm mesh width, dried at 50 C and again forced through a sieve of 1.7 mm mesh width. l-Oxo-2-(2,6-dioxopiperidin-3-yl)-4-amino-isoindoline, the glycine and the saccharin are carefully mixed, the mannitol, the lactose granulate, the stearic acid and the talc are added and the whole is mixed thor-oughly and compressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking groove on the upper side.

Tablets, each containing 10 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-amino-isoindoline, can be prepared in the following manner:

Composition (for 1000 tablets) 1-oxo-2-(2,6-dioxo-piperidin-3-yl)-5-amino-isoindoline .................................... 10.0 g lactose ..........................................328.5 g corn starch ..................................... 17.5 g polyethylene glycol 6000 ............... 5.0 g talc ................................................. 25.0 g magnesium stearate ........................ 4.0 g demineralized water ...................... q.s.

The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
Then the active imide ingredient, lactose, talc, magnesium stearate and half of the starch are intimately mixed. The other half of the starch is suspended in 65 mL of water and this suspension is added to a boiling solution of the polyethylene glycol in 260 mL of water. The resulting paste is added to the pulverulent substances, and the whole is mixed and granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and com-pressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking notch on the upper side.

Gelatin dry-filled capsules, each containing 100 mg of I -oxo-2-(2,6-dioxopip-eridin-3-yl)-6-aminoisoindoline, can be prepared in the following manner:

Composition (for 1000 capsules) I -oxo-2-(2,6-dioxo-piperidin-3-yl)-6-amino-isoindoline ................................... 100.0 g microcry stall ine cellulose ............. 30.0 g sodium lauryl sulfate ...................... 2.0 g magnesium stearate ........................ 8.0 g The sodium lauryl sulfate is sieved into the I -oxo-2-(2,6-dioxopiperidin-3-yl)-6-aminoisoindoline through a sieve of 0.2 mm mesh width and the two components are intimately mixed for 10 minutes. The microcrystalline cellulose is then added through a sieve of 0.9 mm mesh width and the whole is again intimately mixed for 10 minutes. Finally, the magnesium stearate is added through a sieve of 0.8 mm width I

and, after mixing for a further 3 minutes, the mixture is introduced in portions of 140 mg each into size 0 (elongated) gelatin dry-fill capsules.

A 0.2% injection or infusion solution can be prepared, for example, in the following manner:

1-oxo-2-(2,6-dioxo-piperidin-3-yl)-7-amino-isoindoline ..................................... 5.0 g sodium chloride ............................ 22.5 g phosphate buffer pH 7.4 ..............300.0 g demineralized water .............. to 2500.0 mL
1-Oxo-2-(2,6-dioxopiperidin-3-yl)-7-aminoisoindoline is dissolved in 1000 mL of water and filtered through a microfilter. The buffer solution is added and the whole is made up to 2500 mL with water. To prepare dosage unit forms, portions of 1.0 or 2.5 mL each are introduced into glass ampoules (each containing respectively 2.0 or 5.0 mg of imide).

Tablets, each containing 50 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can be prepared in the following manner:

Constituents (for 1000 tablets) 1-oxo-2-(2,6-dioxo-piperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline .....................50.0 g lactose ............................................ 50.7 g wheat starch .................................... 7.5 g polyethylene glycol 6000 ................ 5.0 g talc ................................................... 5.0 g magnesium stearate ......................... 1.8 g demineralized water ....................... q.s.

The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
The active ingredient, lactose, talc, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspen-sion is added to a boiling solution of the polyethylene glycol in 100 mL of water.
The resulting paste is added to the pulverulent substances and the mixture is granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets, each containing 100 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroisoindoline, can be prepared in the following manner:

Constituents (for 1000 tablets) 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroisoindoline ...... 100.0 g lactose ..........................................100.0 g wheat starch .................................. 47.0 g magnesium stearate ........................ 3.0 g All the solid ingredients are first forced through a sieve of 0.6 mm mesh width. The active ingredient, lactose, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspension is added to 100 mL of boiling water. The resulting paste is added to the pulverulent substances and the mixture is granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets for chewing, each containing 75 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can be prepared in the following manner:

T

Composition (for 1000 tablets) 1 -oxo-2-(2,6-dioxo-piperidin-3-yl)- 4,5,6,7-tetra-fluoroisoindoline ........................... 75.0 g mannitol ....................................... 230.0 g lactose ..........................................150.0 g talc ................................................. 21.0 g glycine ...........................................12.5 g stearic acid .....................................10.0 g saccharin ......... .............................1.5 g 5% gelatin solution ......................... q.s.

All the solid ingredients are first forced through a sieve of 0.25 mm mesh width. The mannitol and the lactose are mixed, granulated with the addition of gelatin solution, forced through a sieve of 2 mm mesh width, dried at 50 C and again forced through a sieve of 1.7 mm mesh width. l-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, the glycine and the saccharin are carefully mixed, the mannitol, the lactose granulate, the stearic acid and the talc are added and the whole is mixed thoroughly and compressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking groove on the upper side.

Tablets, each containing 10 mg of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethylisoindoline, can be prepared in the following manner:

Composition (for 1000 tablets) 1-oxo-2-(2,6-dioxo-piperidin-3-yl)- 4,5,6,7-tetramethylisoindoline .................. 10.0 g lactose ..........................................328.5.g corn starch ..................................... 17.5 g polyethylene glycol 6000 ............... 5.0 g talc ..... ........................................ 25.0 g magnesium stearate ........................ 4.0 g demineralized water ...................... q.s.

The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
Then the active imide ingredient, lactose, talc, magnesium stearate and half of the starch are intimately mixed. The other half of the starch is suspended in 65 mL of water and this suspension is added to a boiling solution of the polyethylene glycol in mL of water. The resulting paste is added to the pulverulent substances, and the whole is mixed and granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and com-pressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking notch on the upper side.

Gelatin dry-filled capsules, each containing 100 nlg of 1-oxo-2-(2,6-dioxo-piperidin-3-yl)-4,5,6,7-tetramethoxyisoindoline, can be prepared in the following manner:

Composition (for 1000 capsules) 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxy-isoindoline ................................... 100.0 g microcrystalline cellulose ............. 30.0 g sodium lauryl sulfate ...................... 2.0 g magnesium stearate ........................ 8.0 g The sodium lauryl sulfate is sieved into the 1-oxo-2-(2.6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoindoline through a sieve of 0.2 mm mesh width and the two components are intimately mixed for 10 minutes. The microcrystalline cellulose is then added through a sieve of 0.9 mm mesh width and the whole is again intimately mixed for 10 minutes. Finally, the magnesium stearate is added through a sieve of 0.8 mm width and, after mixing for a further 3 minutes, the mixture is introduced in portions of 140 mg each into size 0 (elongated) gelatin dry-fill capsules.

A 0.2% injection or infusion solution can be prepared, for example, in the following manner:

7? 1 1 I -oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline ........ 5.0 g sodium chloride ............................ 22.5 g phosphate buffer pH 7.4 ..............300.0 g demineralized water .............. to 2500.0 mL
I-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline is dissolved in 1000 mL of water and filtered through a microfilter. The buffer solution is added and the whole is made up to 2500 mL with water. To prepare dosage unit forms, portions of 1.0 or 2.5 mL each are introduced into glass ampoules (each containing respec-tively 2.0 or 5.0 mg of imide).

Tablets, each containing 50 mg of I -oxo-2-(2,6-dioxo-3-methylpiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can be prepared in the following manner:

Constituents (for 1000 tablets) I -oxo-2-(2,6-dioxo-3-methyl piperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline ..................... 50.0 g lactose ............................................50.7 g wheat starch .................................... 7.5 g polyethylene glycol 6000 ................ 5.0 g talc ................................................... 5.0 g magnesium stearate ......................... 1.8 g demineralized water ........................ q.s.

The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
The active ingredient, lactose, talc, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspen-sion is added to a boiling solution of the polyethylene glycol in 100 mL of water. The resulting paste is added to the pulverulent substances and the mixture is granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets, each containing 100 mg of l -oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoiso-indoline, can be prepared in the following manner:

Constituents (for 1000 tablets) 1-oxo-2-(2,6-dioxo-piperidin-3-yl)-4-amino isoindoline ................................... 100.0 g lactose ..........................................100.0 g to wheat starch .................................. 47.0 g magnesium stearate ........................ 3.0 g All the solid ingredients are first forced through a sieve of 0.6 mm mesh width.
The active ingredient, lactose, magnesium stearate and half of the starch then are mixed. The other half of the starch is suspended in 40 mL of water and this suspen-sion is added to 100 mL of boiling water. The resulting paste is added to the pulveru-lent substances and the mixture is granulated, if necessary with the addition of water.
The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 6 mm diameter which are concave on both sides.

Tablets for chewing, each containing 75 mg of 2-(2,6-dioxo-3-methylpiperidin-3-yl)-4-aminophthalimide, can be prepared in the following manner:

Composition (for 1000 tablets) 2-(2,6-dioxo-3-methylpiperidin-3-yl)-4-aminophthalimide ............. 75.0 g mannitol .......................................230.0 g lactose .......................................... 150.0 g talc ................................................. 21.0 g glycine ............................................ 12.5 g stearic acid .....................................10.0 g saccharin .......................................... 1.5 g 5% gelatin solution ......................... q.s.
All the solid ingredients are first forced through a sieve of 0.25 mm mesh width.
The mannitol and the lactose are mixed, granulated with the addition of gelatin solu-n 1 tion, forced through a sieve of 2 mm mesh width, dried at 50 C and again forced through a sieve of 1.7 mm mesh width. 2-(2,6-Dioxo-3-methylpiperidin-3-yl)-4-aminophthalimide, the glycine and the saccharin are carefully mixed, the mannitol, the lactose granulate, the stearic acid and the talc are added and the whole is mixed thoroughly and compressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking groove on the upper side.

Tablets, each containing 10 mg of 2-(2,6-dioxoethylpiperidin-3-yl)-4-aminophthal-imide, can be prepared in the following manner:

Composition (for 1000 tablets) 2-(2,6-dioxoethylpiperidin-3-yl)-4-aminophthalimide ...................... 10.0 g lactose ..........................................328.5 g corn starch ..................................... 17.5 g polyethylene glycol 6000 ............... 5.0 g talc ................................................. 25.0 g magnesium stearate ........................ 4.0 g demineralized water ...................... q.s.
The solid ingredients are first forced through a sieve of 0.6 mm mesh width.
Then the active imide ingredient, lactose, talc, magnesium stearate and half of the starch are intimately mixed. The other half of the starch is suspended in 65 mL of water and this suspension is added to a boiling solution of the polyethylene glycol in 260 mL
of water. The resulting paste is added to the pulverulent substances, and the whole is mixed and granulated, if necessary with the addition of water. The granulate is dried overnight at 35 C, forced through a sieve of 1.2 mm mesh width and compressed to form tablets of approximately 10 mm diameter which are concave on both sides and have a breaking notch on the upper side.

Gelatin dry-filled capsules, each containing 100 mg of 1-oxo-2-(2,6-dioxo-3--methylpiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can be prepared in the following manner:.

Composition (for 1000 capsules) 1-oxo-2-(2,6-dioxo-3-methylpiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline ................... 100.0 g microcrystalline cellulose ............. 30.0 g sodium lauryl sulfate ...................... 2.0 g magnesium stearate ........................ 8.0 g The sodium lauryl sulfate is sieved into the 1-oxo-2-(2,6-dioxo-3-methylpiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline through a sieve of 0.2 mm mesh width and the two components are intimately mixed for 10 minutes. The microcrystalline cellulose is then added through a sieve of 0.9 mm mesh width and the whole is again intimately mixed for 10 minutes. Finally, the magnesium stearate is added through a sieve of 0.8 mm width and, after mixing for a further 3 minutes, the mixture is introduced in portions of 140 mg each into size 0 (elongated) gelatin dry-fill capsules.

A 0.2% injection or infusion solution can be prepared, for example, in the follow-ing manner:

1 -oxo-2-(2,6-dioxo-3-methyl piperidin-3-yl)-4,5,6,7-tetrafluoro isoindoline ..................................... 5.0 g sodium chloride ............................ 22.5 g phosphate buffer pH 7.4 ..............300.09 demineralized water .............. to 2500.0 mL
1-Oxo-2-(2,6-dioxo-3-methylpiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline is dissolved in 1000 mL of water and filtered through a microfilter. The buffer solution is added and the whole is made up to 2500 mL with water. To prepare dosage unit forms, portions of 1.0 or 2.5 mL each are introduced into glass ampoules (each containing respectively 2.0 or 5.0 mg of imide).

TI , T

Claims (12)

1. An optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, having the following structure:

or a pharmaceutically acceptable salt thereof.

2. An optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, having the following structure:

or a pharmaceutically acceptable salt thereof.

3. The optical isomer of claim 1 or 2 or pharmaceutically acceptable salt thereof having an optical purity of > 95%.

4. An optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline.

5. An optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline.

6. The optical isomer of claim 4 or 5 having an optical purity of > 95%.

7. An optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, having the following structure:

or a pharmaceutically acceptable salt thereof.

8. An optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, having the following structure:

or a pharmaceutically acceptable salt thereof.

9. The optical isomer of claim 7 or 8 or pharmaceutically acceptable salt thereof having an optical purity of > 95%.

10. An optical isomer, which is (R)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline.

11. An optical isomer, which is (S)-1,3-dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline.

12. The optical isomer of claim 10 or 11 having an optical purity of > 95%.